Pericardial Fat and Inflammation in HIV Patients and Controls

Methods and Procedures:

Screening Assessments Patients who potentially meet inclusion criteria will be identified
through the Infectious Disease Center and/or the cardiac catheterization laboratory who have
had a coronary angiogram or cardiac CT angiogram, with or without evidence of CAD. Their
records will be reviewed for inclusion and exclusion criteria and informed consent will be
obtained prior to any research related procedures by a member of the research team for this
project.

A screening visit will be done to obtain informed consent. BUN and Creatinine will be done
at screening to ensure the safety of the MRIprocedure.

In HIV-infected subjects who have not had HIV-1 RNA or CD4+ lymphocyte tests obtained in the
last 60 days, these will be done at screening.

Screening visits will last approximately 30-60 minutes.

Entry Visit - MRI The entry visits will occur within 14 days of the screening assessment.
All female patients of child bearing potential will have a negative pregnancy test.

If BUN/Creatinine results are greater than 14 days old, they will be repeated STAT prior to
beginning MRI procedure.

Whole blood will be drawn for biobanking (PBMCs, Plasma and Serum) as well as fasting lipids
will be obtained.

An IV will be started in the MRI laboratory. All participants will undergo an initial
cardiac MRI with contrast for the evaluation of cardiac function, chamber size, resting
perfusion, pericardial adipose volume/mass, myocardial edema and determination of
infarction.

The GE Signa Horizon 1.5T Magnet will be used to obtain all images. Sequences obtained will
include Double IR, Triple IR, T2 Stars, SSFP and delayed gadolinium enhancement imaging.

After the initial CMR on day 1, Feraheme 5 mg/kg IV will be administered. Feraheme (30
mg/mL) is available for intravenous injection in single use vials. Each vial contains 510 mg
of elemental iron in 17 mL.

Feraheme will be administered as an undiluted intravenous injection delivered at a rate of
up to 1 mL/sec (30 mg/sec) through a filtered needle.

Patients will be monitored for 1 hour after administration for signs of allergic reaction.

The total amount of time for the Entry visit will be about 4 hours. Day 2 or 3 - repeat MRI
A second, limited cardiac MRI will be obtained 48-72 hours after injection of Feraheme to
determine the presence of monocyte/macrophages in adipose tissue.

The GE Signa Horizon 1.5T Magnet will be used to obtain all images. Imaging with T2 Stars
and Triple IR sequences.

Data To be Collected:

MRI data to be collected will include:

Left ventricular and right ventricular ejection fractions Left ventricular wall motion Edema
Resting myocardial perfusion Pericardial adipose tissue volume T2 star values of pericardial
fat pre-Feraheme and post-Feraheme Late gadolinium enhancement (scar assessment and
distribution) Valvular assessment Extracardiac assessment - pericardial effusion, pleural
effusion, ascites Vital signs (height, weight, blood pressure, heart rate, BSA) Additional
Data to be collected if available from the clinical record. Left Heart catheterization
Presence or absence of coronary artery disease. The general anatomy, lesion location and
severity, presence, location and type of stents, left ventricular end diastolic pressure,
systemic blood pressure, aortic valve gradient Echocardiogram Ejection fraction Regional
wall motion abnormalities Inducible ischemia if a stress echo was performed Nuclear Stress
study Presence or absence of ischemia or infarction Distribution of perfusion abnormalities.
Ejection fraction TID (transient ischemic dilatation) End Diastolic Volume End Systolic
Volume Medical History (Presence or Absence of ) Sociodemographic information (age,
race/ethnicity, etc) HIV related medical information Myocardial Infarction
Hypercholesterolemia Hypertension Ventricular Tachycardia Stent Placement Bypass Surgery
Diabetes Peripheral Vascular Disease Stroke Smoking (Lifetime Pack-year history and Current
status) Family History of Heart Disease Medications Current use in the last 30 days of
Anti-hypertensive (Beta Blockers, Calcium Channel Blockers, Ace Inhibitors, Angiotensin
Receptor Blockers, Alpha Antagonists) Current use in the last 30 days of Diuretics Lifetime
use of Statins Current use in the last 30 days of Insulin or other anti-diabetic medications
Lifetime use of Antiretroviral Therapy Laboratory Results HIV-1 Viral Load in the last year
Nadir CD4 Count and CD4 count in the last year (include percentages) CD8/CD8 percent in the
last year Troponins ( around time of MI if applicable) Most recent Creatinine within 90 days
of study entry Most recent BUN within 90 days of study entry If subject has diabetes,
hemoglobin A1C or Fructosamine levels in the last year.

Primary endpoints Pericardial adipose tissue volume Change in T2 star value (assessment for
iron deposition as a marker for inflammation) Secondary endpoints Correlation of
inflammatory biomarkers with Pericardial adipose tissue volume and change in T2 star value
Correlation of prior HIV related and Coronary Heart Disease risk factors and events with
inflammatory biomarkers, pericardial adipose tissue volume and change in T2 star value.

Data Analysis Data will be stored in a RedCap data base. Statistical analysis will be
performed using MedCalc. Continuous variables will be analyzed using analysis of Variance
(ANOVA). Non-continuous variables will be analyzed using chi square tests.

Data Monitoring This study will be monitored by a trained coordinator not involved in the
research project after enrollment of the first subject, fifth subject and last subject. The
monitoring plan will include 100% verification of consent process and inclusion and
exclusion criteria. Monitoring for appropriate documentation and collection of study data
will be done on the first three subjects and after completion of the last subjects on all
study participants. Additional monitoring will be conducted based on risk based assessment
of the monitor.

Data Safety Monitoring Board (DSMB) No DSMB will be established for this study. The risks of
MRI and phlebotomy are low and there are no test agents being used that are not Food and
Drug Administration (FDA) approved. The investigators will monitor closely for adverse
events and if an unanticipated number or severity of events occur, the Institutional Review
Board (IRB) will be notified and a DSMB will be convened.

Data Storage and Confidentiality:

Participant medical information will be stored electronically in a Redcap database. Each
subject will have a unique study ID assigned to their data.

The names and medical record numbers of the research participants will be kept under lock
and key and separate from the actual research data in recap. No identifying information will
be present in the research database.

Access to participant medical information contained within this research project will be
restricted to involved investigators and study personnel.

Password protection of data files will be used. Any paper based data pertaining to
individuals will be stored in a locked secure location.

De-identified participant medical record information will be stored for an indefinite period
of time per institutional policies at the University of Cincinnati (UC).

Setting: University of Cincinnati Medical Center, and affiliated MRI facilities

Laboratory Methods and Facilities:

Peripheral blood mononuclear cells, plasma and serum will be obtained and stored for
subsequent measurement of; hsCRP, IL-6, D-dimer, sCD163, CXCL10, flow cytometry for measures
of monocyte activation and additional studies.

DNA will be isolated and may be sent for genetic testing related to inflammation, lipid
disorders and atherosclerosis.

Fasting lipids will be obtained on the day of the cardiac MRI CD4 counts and HIV RNA
quantitation obtained within 60 days of the cardiac MRI will be recorded.

Estimated Period of Time to Complete the Study: 12 months, ending June 30, 2015.

Inclusion Criteria:

- > 18 years of age

- History of HIV disease or no history of HIV

- Prior history of left heart catheterization or CTA indicating the presence or absence
of CAD.

Exclusion Criteria:

- Unable to have a contrast enhanced cardiac MRI, (i.e. ferromagnetic foreign body,
inability to follow verbal instructions, prior history of allergic reactions to
gadolinium, history of nephrogenic systemic fibrosis, severe renal dysfunction GFR<30
ml/min/1.73 m2, or severe claustrophobia/anxiety);

- Chronic kidney disease patient undergoing hemodialysis or peritoneal dialysis

- Allergy to Iron Compounds;

- Pregnant females.